不同富营养培养基对人肠道菌群的体外培养效果比较分析  

Comparative analysis of in vitro cultivation effects of different rich media on human intestinal microbiota

作　　者：路婧 夏雪娟 黄骏楠 陈泫羽 周璐 董庆利[1] LU Jing;XIA Xuejuan;HUANG Junnan;CHEN Xuanyu;ZHOU Lu;DONG Qingli(School of Health Science and Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China)

机构地区：[1]上海理工大学健康科学与工程学院,上海200093

出　　处：《食品与发酵工业》2024年第24期201-211,共11页Food and Fermentation Industries

基　　金：上海市浦江人才计划(23PJ1409600);上海理工大学创新创业训练计划项目(XJ2023328)。

摘　　要：以富含营养的培养基(rich media,RM)为关键组成部分的培养组学技术的兴起为肠道菌群与宿主互作的机制研究提供了重要保障。为了探究不同RM对人肠道菌群的体外培养效果,采用平板计数法以及16S rDNA高通量测序技术分析YCFA培养基(yeast extract-casein hydrolysate-fatty acid medium)、GAM培养基(gifu anaerobic medium)和一种具有专利保护的ZJ培养基对人肠道菌群的体外培养效果。平板计数法显示在不同培养基中培养不同时间(0~7 d)后菌落总数多呈先上升后下降的趋势,经3种培养基培养后肠道菌群的菌落总数的最高值为:ZJ(9.88 lg CFU/mL)>GAM(9.79 lg CFU/mL)>YCFA(9.68 lg CFU/mL)。高通量测序结果显示体外培养后细菌多样性降低,各培养基会选择性富集不同菌株,其中ZJ培养基培养后群落多样性下降最为明显。β-多样性分析结果显示GAM培养基培养后肠道菌群的组成更接近于未培养时的菌群组成。因此,与YCFA和ZJ培养基相比,GAM培养基是一种更优的适合肠道菌群体外培养的RM培养基。The emergence of culturomics,with rich media(RM)as a crucial component,provides a significant foundation for studying the mechanisms of interaction between the gut microbiota and the host.To explore the in vitro cultivation effects of different RMs on the human gut microbiota,the plate counting method and high-throughput 16S rDNA sequencing technology were employed to analyze the cultivation effects of Yeast extract-casein hydrolysate-fatty acid medium(YCFA medium),Gifu anaerobic medium(GAM medium),and a patented ZJ medium on the human gut microbiota.Plate counting results showed that the total colony count of gut microbiota exhibited a trend of initially increasing and then decreasing after cultivation for different durations(0-7 days)in various culture media.The highest colony count values for the gut microbiota after cultivation in three different media were as follows:ZJ(9.88 lg CFU/mL)>GAM(9.79 lg CFU/mL)>YCFA(9.68 lg CFU/mL).The high-throughput sequencing results revealed a decrease in bacterial diversity after in vitro cultivation,with each culture medium selectively enriching different strains.Among them,the ZJ medium exhibited the most significant decrease in community diversity after cultivation.Theβ-diversity analysis indicated that the gut microbiota’s composition after cultivation in the GAM medium was closer to the composition observed in the uncultivated state.Therefore,compared to YCFA and ZJ media,GAM medium is a more optimal RM medium for the in vitro cultivation of the gut microbiota.

关 键 词：肠道菌群 富营养培养基 菌落总数 多样性分析 厌氧培养 

分 类 号：R37[医药卫生—病原生物学]